world-biochem-regenerative-medicine-2019-scientifictracks

Note:

World Biochem 2019 & Regenerative Medicine 2019

Journal of Genetics and Molecular Biology | Volume 3

Page 32

OF EXCELLENCE

IN INTERNATIONAL

MEETINGS

YEARS

March 25-26, 2019 | Amsterdam, Netherlands

BIOCHEMISTRY AND ENZYMOLOGY

World Congress on

TISSUE ENGINEERING AND REGENERATIVE MEDICINE, STEM CELL RESEARCH

Global Conference on

Joint Event on

PROTEOMIC ANALYSIS OF TARGETS OF SER65 PHOSPHORYLATED UBIQUITIN

Julius T Dongdem, Simon Dawson

and

Robert Layfield

University of Nottingham Medical School, UK

ecent studies have highlighted additional levels of complexity in the post-translational modification of pro-

teins by ubiquitin (Ub). For example, Ub itself can be modulated by phosphorylation to act as a second mes-

senger in PINK1-Parkin mediated mitophagy. However, the full physiological significance of Ub phosphorylation

is unclear. Thus, the project sought to catalogue mammalian target proteins modified by covalent attachment

of phosphoSer65-(poly)Ub.

Ub-WT and Ub-Ser65Asp (phosphomimetic mutant) sequences were engineered to allow overexpression of His/

FLAG-tagged proteins in HEK293T cells. An additional Leu73Pro mutation was introduced to stabilise targets

of Ub modification. Endogenous proteins modified by covalent attachment of transfected Ub sequences were

purified by IMAC. Covalent modification was confirmed by western blotting and targets of modification were

identified by LC-MS/MS.

Western blotting affirmed greater target protein modification by Ub-Ser65Asp and Leu73Pro mutants, the lat-

ter an indication of higher resistance to deubiquitination. Proteomic analysis suggested differential modifica-

tion of various target proteins by Ub compared to Ub-Ser65Asp, including endogenous SUMO2. By transfecting

GFP-SUMO2 and its C-terminal–GG deletion mutant, along with phosphomimetic Ub, we confirm that Ub-Ser-

65Asp modifies SUMO2, rather than vice versa. Finally, we confirm that transfected His/FLAG-SUMO2 is modified

by endogenous phosphoSer65-Ub. SUMO2 represents a novel target of Ser65 phosphorylated ubiquitin.

The topics to be covered include targeted drug delivery, tumor therapies, and remote catheter navigation. It will

be shown how iMRI enhances the safety and efficacy of these procedures.

Julius T Dongdem et al., J Genet Mol Biol 2019, Volume 3

Julius T Dongdem is currently a PhD candidate in the area of molecular cell biology and development at the University

of Nottingham, UK. He has completed an MPhil degree in Biochemistry and Molecular Pharmacology at the University

of Nottingham, UK. Julius had earlier obtained a BSc degree in Biochemistry and Chemistry and MPhil in Biochemistry

and Molecular Medicine from the University of Ghana Legon, Ghana. He is a lecturer at the University for Development

Studies, ghana and has more than 10 articles to his credit in reputable Journals. He has also served in editorial board to

more than seven Journals.

BIOGRAPHY